Human vascular endothelial cells are a rich and regulatable source of secretory sphingomyelinase. Implications for early atherogenesis and ceramide-mediated cell signaling.
We recently reported that macrophages and fibroblasts secrete a Zn2+-dependent sphingomyelinase (S-SMase), which, like lysosomal SMase, is a product of the acid SMase gene. S-SMase may cause subendothelial retention and aggregation of lipoproteins during atherogenesis, and the acid SMase gene has been implicated in ceramide-mediated cell signaling, especially involving apoptosis of endothelial cells. Because of the central importance of the endothelium in each of these processes, we now sought to examine the secretion and regulation of S-SMase by vascular endothelial cells. Herein we show that cultured human coronary artery and umbilical vein endothelial cells secrete massive amounts of S-SMase (up to 20-fold more than macrophages). Moreover, whereas S-SMase secreted by macrophages and fibroblasts is almost totally dependent on the addition of exogenous Zn2+, endothelium-derived S-SMase was partially active even in the absence of added Zn2+. Secretion of S-SMase by endothelial cells occurred both apically and basolaterally, suggesting an endothelial contribution to both serum and arterial wall SMase. When endothelial cells were incubated with inflammatory cytokines, such as interleukin-1beta and interferon-gamma, S-SMase secretion by endothelial cells was increased 2-3-fold above the already high level of basal secretion, whereas lysosomal SMase activity was decreased. The mechanism of interleukin-1beta-stimulated secretion appears to be through increased routing of a SMase precursor protein through the secretory pathway. In summary, endothelial cells are a rich and regulatable source of enzymatically active S-SMase, suggesting physiologic and pathophysiologic roles for this enzyme.
Marathe
S
S
Department of Medicine, Columbia University, New York, New York 10032, USA.
Schissel
S L
SL
Yellin
M J
MJ
Beatini
N
N
Mintzer
R
R
Williams
K J
KJ
Tabas
I
I
D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000201:enzymology*; D006801:Humans; D007150:Immunohistochemistry; D007372:Interferons / Q000494:pharmacology; D007378:Interleukins / Q000494:pharmacology; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008247:Lysosomes / Q000201:enzymology; D009131:Muscle, Smooth, Vascular / Q000201:enzymology; D011498:Protein Precursors / Q000378:metabolism; D012333:RNA, Messenger / Q000032:analysis; D013108:Sphingomyelin Phosphodiesterase / Q000378:metabolism*
D007378:Interleukins; D011498:Protein Precursors; D012333:RNA, Messenger; D007372:Interferons; D007770:L-Lactate Dehydrogenase; D013108:Sphingomyelin Phosphodiesterase